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| 1 | One-step generation of myostatin gene knockout sheep via the CRISPR/Cas9 system显示文摘The CRISPR/Cas(clustered regularly interspaced short palindromic repeats/CRISPR-associated)adaptive immune system,which was discovered in bacteria and archaea,can specifically degrade invasive viral and plasmid DNA by base pairing between crRNAs(CRISPR RNAs)and the target DNA[1,2].Recently,the Streptococcus pyogenes type II CRISPR system was shown to be able to perform efficient targeted gene disruption by employing three fundamental components:(1)Cas9 endonuclease. | Hongbing HAN Yonghe MA Tao WANG Ling LIAN Xiuzhi TIAN Rui HU Shoulong DENG Kongpan LI Feng WANG Ning LI Guoshi LIU Yaofeng ZHAO Zhengxing LIAN | 2014 | Frontiers of Agricultural Science and Engineering2014，1，1： | 8 |
| 2 | The Zα domain of fish PKZ facilitates the B-Z conformational transition of oligonucleotide DNAs with d(GC), inserts显示文摘PKZ (象 PKR 一样) 在鱼作为 elF2 kinase 家庭的一个成员被发现,它在 C 终端并且也拥有 eIF2 kinase 的一个保存催化领域在 N 终端的二个 Z-DNA-binding 领域(Z1 和 Z2 ) 。PKZ 能通过 Z 的绑定被激活到 Z-DNA。然而, PKZ Z 的规章的功能仍然保持不清楚。调查 PKZ Z 怎么与 Z-DNA 交往的分子的机制,我们在 Escherichia coli 罗塞达碑紧张表示了 Z 多肽 Z12 并且在 Ni-NTA 上由亲密关系层析净化了树脂。oligonucleotide DNA 与的不同长度各种各样插入,也就是 d (GC ) n (n = 6, 8, 10, 13 ) , d (TA ) n (n = 6, 10 ) , non-d (GC ) ,和 non-d (TA ) ,被设计并且综合。圆形的二色性光谱和胶化活动性移动试金被用来在不同 oligonucleotide DNA 的 conformational 转变上调查 Z12 的效果。结果证明 oligonucleotide DNA 当 Z12 不在时保留了常规 B-DNA 符合构造。与 Z12 滴定的增加的数量, d (GC ) n 被认出并且在某种程度上把符合构造变换成 Z-DNA。与增加重复数字(从 n = 6 到 n = 13 ) , conformational 转变的趋势变得更明显。然而,有 n 插入的 d (TA ) 的 oligonucleotides 的符合构造面对 Z12 有点变化了,并且 Z12 没在有 non-d (GC ) 或 non-d (TA ) 的 oligonucleotides 的转变插入的 conformational 上有效果。胶化活动性移动试金进一步证明 Z12 能与 d (GC ) 绑在 oligonucleotide 10。换句话说, Z12 能与 n 插入到 Z-DNA 符合构造的 d (GC ) 转 oligonucleotides 并且与高亲密关系绑在它。 | Puzhong Lu Shoulong Deng Youlin Zhu Yongbin Yan Yong Liu Chengyu Hu | 2012 | Acta Biochimica et Biophysica Sinica2012，44，11： | 3 |
| 3 | The Zα domain of fish PKZ converts DNA hairpin with d(GC)n inserts to Z-conformation显示文摘PKZ,蛋白质 kinase 包含 Z-DNA 领域,是脊椎动物 eIF2 kinase 家庭的一个新奇成员。在 N 终点在 C 终点和二个 Z-DNA 有约束力的领域(Z1 和 Z2 ) 包含一个催化领域, PKZ 能通过 Z 的绑定被激活到 Z-DNA。然而, PKZ Z 的规章的功能尚待被证实。这里,蛋白质被表示在 DNA conformational 转变,野类型的 Z1Z2 和 11 异种上理解 PKZ Z 的影响并且净化。同时, DNA hairpinsd (GC ) 的几不同长度 nT4 (GC ) n (n = 26 ) 并且 RNA hairpinr (GC ) 6T4 (GC ) 6 被综合。这些合成 DNA 或 RNA 发卡的符合构造上的 Z1Z2 和变异的蛋白质的效果被使用圆形的二色性光谱和胶化活动性移动试金调查。结果证明 DNA 发卡当 Z1Z2 不在时保留了常规 B-DNA 符合构造,当一些 DNA 发卡(n3 ) 在 Z1Z2 正式就职下面被变换成 Z 符合构造时。趋势按比例与 GC 重复的增加的数量被联系。与 Z1Z2 比较, Z1Z1 而非 Z2Z2 在变换 d (GC ) 显示了一个更高的能力 6T4 (GC ) 6 从 B- 到 Z-DNA。这些结果证明那 Z1 子域比 Z2 子域在 BZ conformational 转变的过程起了一个更必要的作用。变异的蛋白质(K34A, N38A, R39A, Y42A, P57A, P58A,和 W60A ) 不能变换 d (GC ) 6T4 (GC ) 6 进 Z-DNA,而 S35A 或 K56A 保留了一些部分活动。有趣地, Z1Z2 也能导致 r (GC ) 6T4 (GC ) 从 A 符合构造的 6 RNA 到在适当条件下面的 Z 符合构造。 | Puzhong Lu Shoulong Deng Chuxin Wu Youlin Zhu Yong Liu Gang Lin Yongbin Yan Chengyu Hu | 2013 | Acta Biochimica et Biophysica Sinica2013，45，12： | 2 |
| 4 | Over-expression of Toll-like receptor 2 up-regulates heme oxygenase-1 expression and decreases oxidative injury in dairy goats显示文摘Background: Mastitis, an infection caused by Gram-positive bacteria, produces udder inflammation and oxidative injury in milk-producing mammals. Toll-like receptor 2(TLR2) is important for host recognition of invading Grampositive microbes. Over-expression of TLR2 in transgenic dairy goats is a useful model for studying various aspects of infection with Gram-positive bacteria, in vivo.Methods: We over-expressed TLR2 in transgenic dairy goats. Pam3CSK4, a component of Gram-positive bacteria,triggered the TLR2 signal pathway by stimulating the monocytes-macrophages from the TLR2-positive transgenic goats, and induced over-expression of activator protein-1(AP-1), phosphatidylinositol 3-kinase(PI3K) and transcription factor nuclear factor kappa B(NF-κB) and inflammation factors downstream of the signal pathway.Results: Compared with wild-type controls, measurements of various oxidative stress-related molecules showed that TLR2, when over-expressed in transgenic goat monocytes-macrophages, resulted in weak lipid damage, high level expression of anti-oxidative stress proteins, and significantly increased m RNA levels of transcription factor NF-E2-related factor-2(Nrf2) and the downstream gene, heme oxygenase-1(HO-1). When Pam3CSK4 was used to stimulate ear tissue in vivo the HO-1 protein of the transgenic goats had a relatively high expression level.Conclusions: The results indicate that the oxidative injury in goats over-expressing TLR2 was reduced following Pam3CSK4 stimulation. The underlying mechanism for this reduction was increased expression of the anti-oxidation gene HO-1 by activation of the Nrf2 signal pathway. | Shoulong Deng Kun Yu Wuqi Jiang Yan Li Sutian Wang Zhuo Deng Yuchang Yao Baolu Zhang Guoshi Liu Yixun Liu Zhengxing Lian | 2017 | Journal of Animal Science and Biotechnology2017，8，2： | 2 |
| 5 | Efficient production of pronuclear embryos in breeding and nonbreeding season for generating transgenic sheep overexpressing TLR4显示文摘Background: Brucella is a zoonotic Gram-negative pathogen that causes abortion and infertility in ruminants and humans. TLR4 is the receptor for LPS which can recognize Brucella and initiate antigen-presenting cell activities that affect both innate and adaptive immunity. Consequently, transgenic sheep over-expressing TLR4 are an suitable model to investigate the effects of TLR4 on preventing Brucellosis. In this study, we generated transgenic sheep overexpressing TLR4 and aimed to evaluate the effects of different seasons(breeding and non-breeding season) on superovulation and the imported exogenous gene on growth.Results: In total of 43 donor ewes and 166 recipient ewes in breeding season, 37 donor ewes and 144 recipient ewes in non-breeding season were selected for super-ovulation and injected embryo transfer to generate transgenic sheep.Our results indicated the no. of embryos recovered of donors and the rate of pronuclear embryos did not show any significant difference between breeding and non-breeding seasons(P > 0.05). The positive rate of exogenous TLR4 tested were 21.21 % and 22.58 % in breeding and non-breeding season by Southern blot. The expression level of TLR4 in the transgenic sheep was 1.5 times higher than in the non-transgenic group(P < 0.05). The lambs overexpressing TLR4 had similar growth performance with non-transgenic lambs, and the blood physiological parameters of transgenic and non-transgenic were both in the normal range and did not show any difference.Conclusions: Here we establish an efficient platform for the production of transgenic sheep by the microinjection of pronuclear embryos during the whole year. The over-expression of TLR4 had no adverse effect on the growth of the sheep. | Yan Li Di Lian Shoulong Deng Xiaosheng Zhang Jinlong Zhang Wenting Li Hai Bai Zhixian Wang Hongping Wu Juncai Fu Hongbing Han Jianzhong Feng Guoshi Liu Ling Lian Zhengxing Lian | 2017 | Journal of Animal Science and Biotechnology2017，8，1： | 2 |
| 6 | Overexpression of Toll-like receptor 4 contributes to the internalization and elimination of Escherichia coli in sheep by enhancing caveolae-dependent endocytosis显示文摘Background:Gram-negative bacterial infections have a major economic impact on both the livestock industry and public health.Toll-like receptor 4(TLR4)plays a crucial role in host defence against Gram-negative bacteria.Exploring the defence mechanism regulated by TLR4 may provide new targets for treatment of inflammation and control of bacterial infections.In a previous study,we generated transgenic sheep overexpressing TLR4 by microinjection to improve disease resistance.The defence mechanism through which TLR4 overexpression protected these sheep against pathogens is still not fully understood.Results:In the present study,we used Escherichia coli to infect monocytes isolated from peripheral blood of the animal model.The overexpression of TLR4 strongly enhanced the percentage of endocytosis and capacity of elimination in monocytes during the early stages of infection.This phenomenon was mainly due to overexpression of TLR4 promoting caveolae-mediated endocytosis.Pretreatment of the transgenic sheep monocytes with inhibitors of TLR4,Src signalling,or the caveolae-mediated endocytosis pathway reduced the internalization of bacteria,weakened the ability of the monocytes to eliminate the bacteria,and increased the pH of the endosomes.Conclusion:Together,our results reveal the effects of TLR4 on the control of E.coli infection in the innate immunity of sheep and provide crucial evidence of the caveolae-mediated endocytosis pathway required for host resistance to invading bacteria in a large animal model,providing theoretical support for breeding disease resistance in the future.Furthermore,Src and caveolin 1(CAV1)could be potentially valuable targets for the control of infectious diseases. | Yao Li Yue Zhao Xueling Xu Rui Zhang Jinlong Zhang Xiaosheng Zhang Yan Li Shoulong Deng Zhengxing Lian | 2021 | Journal of Animal Science and Biotechnology2021，12，4： | 0 |
| 7 | Effect of peroxiredoxin 6 on total and progressive motility of human spermatozoa after cryopreservation显示文摘Sperm cryopreservation is useful in assisted reproductive technology and male fertility preservation.However,freezing and thawing significantly reduces the total and progressive motility of human spermatozoa.In the present study,we explored the effects of peroxiredoxin 6(PRDX6)on total and progressive motility of human spermatozoa after cryopreservation.Semen samples of 20 males with normal parameters were collected and frozen in media supplemented with different concentrations of PRDX6(0 mM,10−5 mM,10−7 mM,and 10−9 mM,respectively).Postthaw total and progressive motility of sperms were measured.The results showed that in comparison with 0 mM,the concentrations of 10−5 mM,10−7 mM,and 10−9 mM of PRDX6 all significantly improved total motility and progressive motility of sperms(p<0.05).The 10−7 mM of PRDX6 showed the best performance.In conclusion,the supplementation of PRDX6 helps to maintain the total and progressive motility of human spermatozoa. | TIECHENG SUN LUYANG CHENG JING MA SHANJIE ZHOU YANDONG ZHANG WENDI ZHOU SHOULONG DENG SHUSONG WANG TIAN LI | 2020 | BIOCELL2020，44，3： | 0 |
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